Sequencing of mRNA, lncRNA, and circRNA in LLC Cells Treated with Ophiopogonin D

Lung cancer, especially non-small cell lung cancer (NSCLC), is the leading cause of cancer-related deaths worldwide. Natural products play a crucial role in drug development, and most current therapies are derived from these ingredients. Ophiopogon D (OP-D) has received widespread attention due to its broad anti-cancer properties, including the ability to inhibit cell proliferation and induce cell apoptosis. In this study, we performed whole transcriptome sequencing on LLC cells treated with OP-D. Cultivate LLC cells using standard methods and use TRIzol ® Extract total RNA using reagents. Evaluate RNA quality using Agilent 5300 Bioanalyzer and quantify using NanoDrop ND-2000. Construct sequencing libraries using only high-quality RNA samples (OD260/280=1.8-2.2, OD260/230 ≥ 2.0, RQN ≥ 6.5, 28S: 18S ≥ 1.0,>1 μ g). We used Illumina Stranded Total RNA Prep and Ribo Zero Plus for library preparation, generating ribosomal RNA removal libraries including mRNA, lncRNA, and circRNA information. These libraries were sequenced on the Illumina NovaSeq Xplus platform at Majorbio in Shanghai, and a large dataset was uploaded in FASTQ format. The main findings indicate that OP-D processing resulted in significant inter group differentiation as shown in principal component analysis (PCA). Differential expression analysis showed significant downregulation of lncMALAT1, confirming MALAT1 as a significant lncRNA. In addition, mRNA analysis showed that OP-D treatment inhibited key features of malignant tumors, including metastasis and proliferation, highlighting its extensive inhibitory effect on carcinogenic processes.